摘要
目的:观察以兔骨髓基质干细胞为种子细胞,珊瑚羟基磷灰石人工骨为载体的组织工程化珊瑚羟基磷灰石人工骨修复骨缺损的效果。方法:实验于2004-06/12在广州军区广州总医院动物实验中心完成。选取成年新西兰大白兔24只,随机分为3组,自体骨组,珊瑚人工骨组,珊瑚人工骨-细胞复合体组,每组8只。采集兔骨髓进行基质干细胞分离、体外培养、诱导扩增、同时制备珊瑚羟基磷灰石人工骨,并使之与兔骨髓基质干细胞复合,制备细胞-珊瑚人工复合体。麻醉后无菌制备兔桡骨15mm骨缺损模型,将自体骨,珊瑚人工骨,珊瑚人工骨-细胞复合体分别埋入到骨缺损处,术后2,4,6,8周对动物进行精神状况、伤口和X射线观察。每次观察后随机选择处死2只动物并取材,以组织学方法对成骨情况进行观察。成骨情况按照Lane-SandhuX射线评分标准进行评价。评价内容包括骨形成(0分无骨形成;1分骨形成占缺损25%;2分骨形成占缺损50%;3分骨形成占缺损75%;4分骨形成充满缺损)、骨连接(0分骨折线清楚;2分骨折线部分存在;4分骨折线消失)、骨塑形(0分未见骨塑形;2分骨髓腔形成;4分皮质骨塑形)3个方面11项,采用SSPS8.0统计软件对其分值进行统计处理。结果:24只实验动物均进入结果分析①兔骨髓基质干细胞培养后与珊瑚人工骨贴附能力:体外培养3d时,骨髓基质干细胞已充分贴附包裹于珊瑚孔隙周边,5d与7d时无明显变化。②成骨情况:在手术后2、4、6、8周细胞-珊瑚人工骨复合体组Lane-SundhuX射线评分明显优于自体骨组[(5.4±0.95,8.7±0.51,9.2±2.81,11.2±1.15);(0.7±0.51,2.4±0.96,4.3±0.94,7.2±1.06),P<0.05];明显优于珊瑚人工骨组[(5.4±0.95,8.7±0.51,9.2±2.81,11.2±1.15);(0.3±0.41,1.3±0.43,1.8±0.42,4.5±1.16);P<0.05。在手术后4,6,8周时,自体骨组Lane-SundhuX射线评分优于珊瑚人工骨组(P<0.05)。③各组不同时间炎症细胞数量:细胞-珊瑚�
AIM:To observe the outcomes of tissue- engineered coralline hydroxyapatite(C HA) in repair bone defect by using rabbit bone marrow stromal cells(BMSCs) as se ed cells and using CHA as the carrier. METHODS:The experiment was completed in the Experiment Center for Animals,Gen eral Hospital of Guangzhou Military Area Command from June to December 2004.Twen ty- four New Zealand big rabbits were randomly and equally divided into 3 group s:autograft group,CHA group and CHA- BMSCs complex group.After anesthesia under aseptic operation,a model of 15- mm bone defect of rabbit radius was made.The stromal cells derived from rabbit bone marrow were cultured and induced to proli ferate in vitro to combine with the synchronously prepared CHA so as to compose CHA- BMSCs complex.After the autologous bone,CHA and the CHA- BMSCs complex we re implanted into the defects of the corresponding groups respectively,the exami nation of mental status and wound and the X- ray observation were performed at 2,4,6 and 8 weeks.Two animals were scarified after each observation interval to harvest the specimens separately after operation.The formation of new bone was o bserved histologically.Lane- Sandhu's X- ray scoring standard was used to grad e the ostogenesis,including bone formation(0 means no bone formation,1 point mea ns 25% bone formation,2 points mean 50% ,3 points mean 75% and 4 points mean bone defect filled with new bone),synostosis(0 means clear fracture line,2 poin ts mean partial fracture line and 4 points mean disappearing fracture line),bone modeling(0 means no bone modeling,2 points mean meduallary cavity and 4 points mean cortical bone modeling).The data were analyzed with SPSS 8.0 statistical so ftware. RESULTS:All the 24 rabbits were included into the result analysis.① The adhe sive ability of bMSCs onto the CHA:After 3- day culture in vitro,the BMSCs had adhered onto the peripheral CHA fully;however,there was no obvious change on the 5th and 7th days.② The formation of new bone:The scores of Lane- Sandhu X- r ay at 2,4
出处
《中国临床康复》
CSCD
北大核心
2005年第18期64-66,i002,共4页
Chinese Journal of Clinical Rehabilitation
基金
广东省医学科研基金(B2004100)~~
